Cited by 184Receptor Mechanisms and Signalingbioluminescence and chemiluminescence researchChemokine receptors and signaling

High-Throughput Screening of G Protein-Coupled Receptor Antagonists Using a Bioluminescence Resonance Energy Transfer 1-Based β-Arrestin2 Recruitment Assay

Open access
Hamdan, Fadi F., Audet, Martin, Garneau, Philippe, Pelletier, Jerry, Bouvier, Michel·2005·SLAS Discovery

Abstract

In this study, the authors developed HEK293 cell lines that stably coexpressed optimal amounts of beta-arrestin2-Rluc and VENUS fusions of G protein-coupled receptors (GPCRs) belonging to both class A and class B receptors, which include receptors that interact transiently or stably with beta-arrestins. This allowed the use of a bioluminescence resonance energy transfer (BRET) 1- beta-arrestin2 translocation assay to quantify receptor activation or inhibition. One of the developed cell lines coexpressing CCR5-VENUS and beta-arrestin2- Renilla luciferase was then used for high-throughput screening (HTS) for antagonists of the chemokine receptor CCR5, the primary co-receptor for HIV. A total of 26,000 compounds were screened for inhibition of the agonist-promoted beta-arrestin2 recruitment to CCR5, and 12 compounds were found to specifically inhibit the agonist-induced beta-arrestin2 recruitment to CCR5. Three of the potential hits were further tested using other functional assays, and their abilities to inhibit CCR5 agonist-promoted signaling were confirmed. This is the 1st study describing a BRET1-beta-arrestin recruitment assay in stable mammalian cells and its successful application in HTS for GPCRs antagonists.

Integrity signals

Registry

Crossref · 10.1177/1087057105275344

Cites

59 of 59 references verified0 citation errors · 0 not checkable

Statistics

Recomputing p-values needs the paper’s full text.

Claims the paper makes

We couldn’t retrieve this paper’s full text, so its claims and self-check weren’t run. The registry and citation checks still apply.

Cites

Flagged first, then by citations

1 of the 60 references couldn’t be matched to a known paper.

Cited by

Sorted by citations

Authors
Hamdan, F. F., Audet, M., Garneau, P., Pelletier, J., & Bouvier, M.
Affiliations
  • Université de Montréal (CA)
  • McGill University (CA)
Venue
SLAS Discovery
Publisher
Elsevier BV
Published
2005-08
Volume
10
Issue
5
Pages
463-475
ISSN
2472-5552 (ISSN-L)
OpenAlex
W2094714066
Cited by
183
References
60
Citations by year
2026: 2 · 2025: 4 · 2024: 2 · 2023: 3 · 2022: 4 · 2021: 9 · 2020: 8 · 2019: 6 · 2018: 8 · 2017: 5 · 2016: 6 · 2015: 17 · 2014: 8 · 2013: 8 · 2012: 16
FWCI
4.64
Type
journal article
Language
en
Open access
Locations
  • SLAS DISCOVERY (publishedVersion, cc-by-nc-nd)
  • PubMed (publishedVersion)
Topics
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Receptor Mechanisms and Signaling
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › bioluminescence and chemiluminescence research
  • Health Sciences › Medicine › Oncology › Chemokine receptors and signaling
Keywords
G protein-coupled receptors, β-arrestin2, CCR5, chemokine receptor antagonists, high-throughput screening, HIV co-receptor, HEK293 cells, receptor antagonists
MeSH terms
beta-Arrestins, Animals, Automation, Calcium, Cell Line, Dose-Response Relationship, Drug, Drug Evaluation, Preclinical, Energy Transfer, HIV, Humans, Luminescent Measurements, Microscopy, Fluorescence, Plasmids, Time Factors, Genes, Reporter, Arrestins, Receptors, CCR5, Protein Transport, Receptors, G-Protein-Coupled, Macromolecular Substances, Renilla, Luciferases, Renilla, Green Fluorescent Proteins

About these checks

Registry, retraction, Cites and statistics results are computed, not judged, with one exception: a printed reference that no index lists is read once by a model before it counts as a citation error. Claims are extracted by a model and kept only when their quote appears verbatim in the text; each cited source is then checked, with a model’s help, for whether it says what the claim relies on. Those verdicts can be wrong — read the claim’s quote and the cited source before relying on one.

Statistics and arithmetic are recomputed, not judged.

Confidence bands are shown only for fields calibrated on our annotated test set. None is calibrated yet, so no band is shown.

What counts as a claim ›How we measure confidence ›How we check this ›

Are you an author, or do you see a problem with this page? Request a review